The Effect of Antigen Dose and Antigen Presenting Process on T Cell Stimulation: A Method for Enrichment of TB10.4 Antigen-specific T-cell Clones

• Published in: Iranian journal of allergy, asthma, and immunology Vol. 20; no. 3; p. 364
• Main Authors: Motiee, Mahdieh, Zavaran Hosseini, Ahmad, Soudi, Sara, Hassanzadeh, Seyed Mehdi
• Format: Journal Article
• Language: English
• Published: Iran Tehran University of Medical Sciences 06.06.2021
• Subjects: Administration, Inhalation; Animal models; Animals; Antigen-presenting cells; Antigens; Antigens, Bacterial - administration & dosage; Antigens, Bacterial - immunology; BCG Vaccine - administration & dosage; BCG Vaccine - immunology; Cell culture; Cell division; Cell Proliferation; Cell therapy; Cell viability; Cell- and tissue-based therapy; Cells, Cultured; Clone cells; Coculture Techniques; Female; Immunotherapy; Immunotherapy, Adoptive; Interferon-gamma - metabolism; Lymph nodes; Lymphocyte Activation; Lymphocytes; Lymphocytes T; Mice; Mice, Inbred BALB C; T-lymphocytes; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; T-Lymphocytes - transplantation; Time Factors; γ-Interferon; Antigens; Clone cells; T-lymphocytes; Antigen-presenting cells; Immunotherapy; Cell- and tissue-based therapy
• ISSN: 1735-1502; 1735-5249
• DOI: 10.18502/ijaai.v20i3.6338

T-lymphocytes have critical functions in the immune responses against viral and intracellular bacterial infections as well as cancers. Antigen (Ag)-specific T-lymphocyte clones enriched and expanded in vitro are valuable tools in the study of immune responses in animal models and adoptive T-cell therapy of patients with cancer or infection. We described a method for inducing, enriching, and replicating Ag-specific poly-clonal T-cells from BALB/c mice infected with live Bacillus Calmette Guérin (BCG) bacterium. During a 7-8 days procedure, T-lymphocytes were purified from immune cells of lymph nodes stimulated with immunodominant Ag of BCG, TB10.4, and expanded by interleukin -2 cytokine. We evaluated the effect of Ag doses (1, 10, and 100 µg/mL) and exposure method of Ag presenting cells (APCs) to T-cells, on T-cells’ proliferation, viability, and Interferon-gamma (IFN-γ) secretion at 2, 5, and 7 days after Ag stimulation. Increasing Ag concentration increased the average cell division, but at the highest dose of Ag (100 µg/mL), T-cell viability is decreased. Only clones induced by 10 µg/mL Ag produced a desirable amount of IFN-γ. Incubation of Ag and APCs, 24 h before T-lymphocytes addition, increased the proliferation and viability of cells. T cells are in a more favorable condition around day 5 of Ag stimulation in terms of proliferation and survival, and it is the desired time for T cell restimulation. For optimal preparation of specific T-cells for adoptive cell transfer, optimization of Ag dose, the order of APCs and T-cells exposure with Ag, and the duration of initial Ag stimulation, as well as the time for restimulation, is essential.